LOCUS       sequence-209249-        6936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector with hSynapsin promoter, Cre-OFF hCAR (for efficient 
            CAV-2 infection) and Cre-ON ChETA (for optogenetic activation).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6936)
  AUTHORS   Li SJ, Vaughan A, Sturgill JF, Kepecs A
  TITLE     A Viral Receptor Complementation Strategy to Overcome CAV-2 Tropism 
            for Efficient Retrograde Targeting of Neurons.
  JOURNAL   Neuron. 2018 Jun 6;98(5):905-917.e5. doi: 
            10.1016/j.neuron.2018.05.028.
  PUBMED    29879392
REFERENCE   2  (bases 1 to 6936)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.neuron.2018.05"; journalName: "Neuron"; date: "2018-06-6-
            6"; volume: "98"; issue: "5"; pages: "905-917.e5"
COMMENT     Sequence Label: pAAV-hSyn-DIO {hCAR}off-{ChETA}on-WPRE
FEATURES             Location/Qualifiers
     source          1..6936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   2..142
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   13..142
                     /label=AAV2 ITR
     promoter        187..634
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(672..705)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(756..789)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      795..804
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1896..1925
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(1940..1966)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             complement(1940..1966)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(1985..2911)
                     /codon_start=1
                     /product="H134R mutant of an N-terminal fragment of 
                     channelrhodopsin-2 from the alga Chlamydomonas reinhardtii 
                     (Nagel et al., 2005)"
                     /label=ChR2(H134R)
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYATWLLTCPVILIRLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     regulatory      2908..2917
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2922..2955
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    3006..3039
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3064..3652
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3117..3137)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3535..3546)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3684..4160
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3883..3902)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4200..4340
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4200..4329
                     /label=AAV2 ITR
     rep_origin      4415..4870
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4502..4521)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4712..4733
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4887..4906)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5006..5028
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5066..5084)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5152..5256
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5257..5325,5326..6117)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5475..5494)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6288..6876
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6777..6796
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt
       61 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
      121 aactccatca ctaggggttc ctgcggccgc acgcgtgtgt ctagactgca gagggccctg
      181 cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg ggtgggggtg cctacctgac
      241 gaccgacccc gacccactgg acaagcaccc aacccccatt ccccaaattg cgcatcccct
      301 atcagagagg gggaggggaa acaggatgcg gcgaggcgcg tgcgcactgc cagcttcagc
      361 accgcggaca gtgccttcgc ccccgcctgg cggcgcgcgc caccgccgcc tcagcactga
      421 aggcgcgctg acgtcactcg ccggtccccc gcaaactccc cttcccggcc accttggtcg
      481 cgtccgcgcc gccgccggcc cagccggacc gcaccacgcg aggcgcgaga taggggggca
      541 cgggcgcgac catctgcgct gcggcgccgg cgactcagcg ctgcctcagt ctgcggtggg
      601 cagcggagga gtcgtgtcgt gcctgagagc gcagtcgaga aggtaccgga tcctctagag
      661 tcgactccgg aataacttcg tataggatac tttatacgaa gttatgcaga atggtagctg
      721 gattgtagct gctattagca atatgaaacc tcttaataac ttcgtatagc atacattata
      781 cgaagttatg gcgcgccacc atggcgctcc tgctgtgctt cgtgctcctg tgcggagtag
      841 tggatttcgc cagaagtttg agtatcacta ctcctgaaga gatgattgaa aaagccaaag
      901 gggaaactgc ctatctgccg tgcaaattta cgcttagtcc cgaagaccag ggaccgctgg
      961 acatcgagtg gctgatatca ccagctgata atcagaaggt ggatcaagtg attattttat
     1021 attctggaga caaaatttat gatgactact atccagatct gaaaggccga gtacatttta
     1081 cgagtaatga tctcaaatct ggtgatgcat caataaatgt aacgaattta caactgtcag
     1141 atattggcac atatcagtgc aaagtgaaaa aagctcctgg tgttgcaaat aagaagattc
     1201 atctggtagt tcttgttaag ccttcaggtg cgagatgtta cgttgatgga tctgaagaaa
     1261 ttggaagtga ctttaagata aaatgtgaac caaaagaagg ttcacttcca ttacagtatg
     1321 agtggcaaaa attgtctgac tcacagaaaa tgcccacttc atggttagca gaaatgactt
     1381 catctgttat atctgtaaaa aatgcctctt ctgagtactc tgggacatac agctgtacag
     1441 tcagaaacag agtgggctct gatcagtgcc tgttgcgtct aaacgttgtc cctccttcaa
     1501 ataaagctgg actaattgca ggagccatta taggaacttt gcttgctcta gcgctcattg
     1561 gtcttatcat cttttgctgt cgtaaaaagc gcagagaaga aaaatatgaa aaggaagttc
     1621 atcacgatat cagggaagat gtgccacctc caaagagccg tacgtccact gccagaagct
     1681 acatcggcag taatcattca tccctggggt ccatgtctcc ttccaacatg gaaggatatt
     1741 ccaagactca gtataaccaa gtaccaagtg aagactttga acgcactcct cagagtccga
     1801 ctctcccacc tgctaaggta gctgccccta atctaagtcg aatgggtgcg attcctgtga
     1861 tgattccagc acagagcaag gatgggtcta tagtagagca gaagctgatc agcgaggagg
     1921 acctgtaatt aattaattag gcgtagtcgg gcacgtcgta ggggtacatg gtggcggccg
     1981 ctggcacggc tccggcctcg gcttcgtctt cgacgagagt ctcgacctcg atctccgttc
     2041 cgccgatgtt cagtttggtg gttttgcgga tatctccgtg aatcaatatg tgctcgtgga
     2101 tcaggacgcg caggtagtgt cccaacaacc cccaacaatt tttactcatc agatcaataa
     2161 tcgtgtgacc tacggtggag ccatagacgc tcaggacgcc aaaaccttcg ggccccaaaa
     2221 tgaagagaat tgggaacata ccccagctca cgaaaaacag ccatgccatg ccggtcacga
     2281 cctggcggca ccgacccttt ggcacagtat gataaccctc gatatatgct ttggcggcgt
     2341 gaaaaaatgt gttcgcgcca tagcacaatc caagacaaaa gaagatgact ttaacatagc
     2401 cggttgccat ggcgctggta gccccccaca cgatagtccc gatgtctgag acaaggagtc
     2461 ccatggttct cctgctgtag tcgttgctca ggccggtgag gttgctcagg cggataagga
     2521 tgacaggaca agtgagcagc caggttgcat agcgcagcca ctgcacccgg tgtcctgtgg
     2581 caaggtagag catagaggga ttcttaaact caaaaaagaa ctcgagaatc accttaacca
     2641 tttcaatggc gcacacatag atctcctccc agccgcatgt agatttccag gtttggtagg
     2701 catagaacat cagcagcaaa atgctgaatc ctgctgcaag ccactgcagg acatttgacg
     2761 cggtctgagc gccgttcgtg ccgcgagatt caatccatcc ggcacagtaa cattgatcct
     2821 cagggaccag gacggacccg ttcaccacca caggattagt aacgaacaaa agttcgcgtc
     2881 cgacggcaga caaagcgccg ccatagtcca tggtggctag cataacttcg tataaagtat
     2941 cctatacgaa gttatttgcc ttaacccaga aattatcact gttattcttt agaatggtgc
     3001 aaagaataac ttcgtataat gtatgctata cgaagttatg aattcgatat caagcttatc
     3061 gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     3121 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     3181 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     3241 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     3301 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     3361 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     3421 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     3481 ctcgcctatg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     3541 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     3601 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac
     3661 cgagcgctgc tcgagagatc tacgggtggc atccctgtga cccctcccca gtgcctctcc
     3721 tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca
     3781 tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga
     3841 gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct
     3901 ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc
     3961 tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat
     4021 ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct
     4081 aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca
     4141 ctgctccctt ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg agcggccgca
     4201 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     4261 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     4321 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     4381 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     4441 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4501 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4561 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4621 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4681 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4741 ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     4801 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     4861 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     4921 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     4981 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     5041 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     5101 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5161 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5221 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5281 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5341 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5401 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5461 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5521 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5581 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5641 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5701 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5761 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     5821 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5881 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5941 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6001 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6061 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6121 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6181 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6241 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6301 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6361 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6421 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6481 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6541 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6601 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6661 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6721 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6781 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6841 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6901 cggttcctgg ccttttgctg gccttttgct cacatg
//
